Regulation of eukaryotic gene expression by transcriptional attenuation.

Wright, Stephanie C.

doi:10.1091/mbc.4.7.661

Cited by 41 publications

(25 citation statements)

References 114 publications

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“…This process, referred to as abortive elongation or premature termination has been observed in various transcription systems (1)(2)(3)(4)(5), as well as in our study of Drosophila RNA polymerase II transcription (6,7). We observed that two distinct classes of complexes were formed after initiation.…”

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confidence: 65%

Unusual Nucleic Acid Binding Properties of Factor 2, an RNA Polymerase II Transcript Release Factor

Zhi¹,

Price

1998

Journal of Biological Chemistry

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Drosophila factor 2, an RNA polymerase II transcript release factor, exhibits a DNA-dependent ATPase activity (Xie, Z., and Price D. H. (1997) J. Biol. Chem. 272, 31902-31907). We examined the nucleic acid requirement and found that only double-stranded DNA (dsDNA) effectively activated the ATPase. Single-stranded DNA (ssDNA) not only failed to activate the ATPase, but suppressed the dsDNA-dependent ATPase. Gel mobility shift assays showed that factor 2 formed stable complexes with dsDNA or ssDNA in the absence of ATP. However, in the presence of ATP, the interaction of factor 2 with dsDNA was destabilized, while the ssDNAfactor 2 complexes were not affected. The interaction of factor 2 with dsDNA was sensitive to increasing salt concentrations and was competed by ssDNA. In both cases, loss of binding of factor 2 to dsDNA was mirrored by a decrease in ATPase and transcript release activity, suggesting that the interaction of factor 2 with dsDNA is important in coupling the ATPase with the transcript release activity. Although the properties of factor 2 suggested that it might have helicase activity, we were unable to detect any DNA unwinding activity associated with factor 2.A common mechanism employed to control the potential of RNA polymerase II to synthesize full-length transcripts is through an early elongation block. This process, referred to as abortive elongation or premature termination has been observed in various transcription systems (1-5), as well as in our study of Drosophila RNA polymerase II transcription (6, 7). We observed that two distinct classes of complexes were formed after initiation. The predominant class undergoes abortive elongation which only gives rise to short transcripts, whereas the second class overcomes early blocks and carries out productive elongation. Based on our results, a model has been proposed for the control of elongation by RNA polymerase II which highlights the function of both negative and positive factors (6, 7). According to the model, the action of negative transcription elongation factors (N-TEF) 1 was responsible for the abortive elongation. The transition from abortive elongation to productive elongation was mediated by the action of positive transcription elongation factors (P-TEF). P-TEFb, one of the components of P-TEF, has recently been purified and identified as a 5,6-dichloro-1-␤-D-ribofuranosylbenzimidazole-sensitive kinase that can phosphorylate the C-terminal domain (CTD) of the largest subunit of RNA polymerase II (8, 9) and is required for Tat-transactivation of transcription from the HIV-LTR (10, 11).One of the components of N-TEF, factor 2, was originally identified due to its ability to suppress the appearance of shorter than full-length transcripts during transcription in vitro (12). Factor 2 is a 154-kDa protein that associates stably with early elongation complexes under low salt conditions, but dissociates in 1 M KCl (13). Factor 2 causes the release of transcripts by RNA polymerase II in an ATP-dependent manner and results in premature termina...

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confidence: 65%

Unusual Nucleic Acid Binding Properties of Factor 2, an RNA Polymerase II Transcript Release Factor

Zhi¹,

Price

1998

Journal of Biological Chemistry

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“…Thus, its activity seemed to be restricted to mRNA elongation. Transcriptional elongation or antitermination is an important element of the regulation of gene expression in prokaryotic, eukaryotic, and viral systems (16).…”

Section: Discussionmentioning

confidence: 99%

Transcription elongation factor of respiratory syncytial virus, a nonsegmented negative-strand RNA virus.

Collins

Hill

Cristina

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

261

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RNA synthesis by the paramyxovirus respiratory syncytial virus, a ubiquitous human pathogen, was found to be more complex than previously appreciated for the nonsegmented negative-strand RNA viruses. Intracellular RNA replication of a plasmid-encoded "minigenome" analog of viral genomic RNA was directed by coexpression of the N, P, and L proteins. But, under these conditions, the greater part of mRNA synthesis terminated prematurely. This difference in processivity between the replicase and the transcriptase was unanticipated because the two enzymes ostensively shared the same protein subunits and template. Coexpression of the M2 gene at a low level of input plasmid resulted in the efficient production of full-length mRNA and, in the case of a dicistronic minigenome, sequential transcription. At a higher level, coexpression of the M2 gene inhibited transcription and RNA replication. The M2 mRNA contains two overlapping translational open reading frames (ORFs), which were segregated for further analysis. Expression of the upstream ORFI, which encoded the previously described 22-kDa M2 protein, was associated with transcription elongation. A model involving this protein in the balance between transcription and replication is proposed. ORF2, which lacks an assigned protein, was associated with inhibition of RNA synthesis. We propose that this activity renders nucleocapsids synthetically quiescent prior to incorporation into virions.The paramyxovirus human respiratory syncytial virus (RSV) is a nonsegmented negative-strand RNA virus (1). Information on the molecular biology of this large group of viruses is based mainly on the rhabdovirus vesicular stomatitis virus and the paramyxovirus Sendai virus (2-5). The RNA genomes of these prototype viruses are tightly encapsidated by the major nucleocapsid N or NP protein and also are associated with the nucleocapsid phosphoprotein P and the large L polymerase protein. Transcription initiates at the 3' extragenic leader region of genomic RNA and proceeds by a sequential, polar, stop-start mechanism that produces subgenomic mRNAs. RNA replication involves a switch to readthrough synthesis to produce a genome-length positive strand replicative intermediate (antigenome), which also is tightly encapsidated and serves as the template for production of progeny genomes.The genome of RSV is 15,222 nt long. It encodes 10 major species of mRNA and 10 major viral proteins, compared with 5-7 mRNAs for the prototype nonsegmented negative-strand RNA viruses. Several RSV proteins lack known counterparts in most or all other nonsegmented negative-strand virusesnamely, two The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.in the middle of the molecule (see Fig. 4); the upstream ORF1 encodes the M2(ORF1) protein and the downstream ORF2 lacks an assigned protein (6). Other RSV proteins, such as N, P, and L, appear to have dire...

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“…In procaryotes transcriptional attenuation or pausing results from different mechanisms, involving specific D N A sequences, protein-mediated inhibition of RNA polymerase, or specific secondary structures adopted by the nascent transcript (Das, 1993). In eucaryotes examples of the control of gene expression at the level of elongation have been described for many genes (Wright, 1993), such as the hsp70 gene in Drosophila (O'Brien and Lis, 1991) or the c-myc gene in mammals (Krumm e t a]., 1995). Very little is known o n the regulation of elongation during rDNA transcription.…”

Section: Discussionmentioning

confidence: 99%

Two Ribosomal DNA‐Binding Factors Interact with a Cluster of Motifs on the 5′ External Transcribed Spacer, Upstream from the Primary Pre‐rRNA Processing Site in a Higher Plant

Caparrós-Ruiz

Lahmy

Piersanti

et al. 1997

European Journal of Biochemistry

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In radish the primary processing site in pre-rRNA has been mapped to a TTTTCGCGC sequence Nuclear genes coding for the 25S, 5.8s and 18s rRNAs are organised as head-to-tail tandem repeats of the rRNA sequences separated by two short spacers and a large intergenic spacer (IGS). Transcription of this unit by RNA polymerase I gives a primary rRNA precursor (pre-rRNA) comprising the rRNA and transcribed spacer sequences. Pre-rRNA processing results in removal of spacer sequences by a combination of endonucleolytic cleavages at specific sites, exonucleolytic RNA degradation, and base modification of rRNAs. All these events occur in the nucleolus (Sollner-Webb and Mougey, 1991 ;Moss and Stefanovsky, 1995).In eucaryotes, ribosomal DNA (rDNA) transcription starts at a major transcription initiation site (TIS) located in the IGS and proceeds through the 5' external transcribed spacer (5' ETS). The first pre-rRNA-processing event occurs in the nascent transcript and corresponds to an endonucleolytic cleavage at a speCorrespondence to M. Echevem'a,

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Regulation of eukaryotic gene expression by transcriptional attenuation.

Cited by 41 publications

References 114 publications

Unusual Nucleic Acid Binding Properties of Factor 2, an RNA Polymerase II Transcript Release Factor

Unusual Nucleic Acid Binding Properties of Factor 2, an RNA Polymerase II Transcript Release Factor

Transcription elongation factor of respiratory syncytial virus, a nonsegmented negative-strand RNA virus.

Two Ribosomal DNA‐Binding Factors Interact with a Cluster of Motifs on the 5′ External Transcribed Spacer, Upstream from the Primary Pre‐rRNA Processing Site in a Higher Plant

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